糖尿病前期患者循环miR-103b表达、临床意义及靶基因功能研究

摘要

AIM:To investigate whether serum microRNA(miR)-103b plays a critical role in the pathogene-sis of type 2 diabetes mellitus(T2DM)and pre-diabetic syndrome.METHODS:Bioinformatic analysis was used for iden-tification of miR-103b and its targets,and the results were assessed by real-time PCR and receiver-operating characteristic (ROC)curve analysis in 48 patients with pre-diabetes mellitus(pre-DM),47 patients with noncomplicated diabetes melli-tus(NCDM),and 50 healthy individuals.RESULTS:miR-103b was significantly down-regulated in serum from the pa-tients with pre-DM and NCDM compared with healthy individuals.The ROC curve analysis found that the area under the curve(AUC)of miR-103b was 0.887(95% CI 0.809~0.944).The bioinformatic analysis has demonstrated that miR-103b has a high degree of site conservation among different mammalian species,such as Homo sapiens,Mus musculus,Rat-tus norvegicus,Pongo pygmaeus,Sus scrofa,etc.Fifty-three potential targets of miR-103b were predicted, most of which were involved in MAPK,Wnt,insulin and Ras signaling pathways,and enriched in various biological processes(such as phosphoprotein,DNA regulation transcription,cell growth and proliferation,apoptosis, cell cycle, etc), molecular func-tions(such as protein binding)and cell component(such as filamentous actin).CONCLUSION:Serum miR-103b can be used as an objective complement to traditional diagnosis of pre-diabetes,indicating important implications regarding the distinguish of the undiagnosed cases between diabetes and pre-diabetes by circulating miRNA.%目的:探讨2型糖尿病前期患者循环血清中微小RNA(miR)-103b的表达变化与临床意义,并分析其靶基因生物信息学功能.方法:收集糖尿病前期患者(n=48)、糖尿病无并发症患者(n=47)及正常健康人群(n=50)的血清标本,用real-time PCR检测各血清样本中miR-103b的表达水平,并比较3组miR-103b表达水平的差异与临床病理特征之间的相关性,运用受试者工作特征(ROC)曲线评估诊断效率及特异性.利用生物信息学技术,对hsa-miR-103b的特征、进化保守性、靶基因功能以及GO信号通路等进行深入分析.结果:Real-time PCR结果显示,与健康人群相比,循环血清中miR-103b在糖尿病前期及糖尿病无并发症患者的表达显著降低(P<0.05).ROC曲线分析结果显示,糖尿病前期患者血清中miR-103b的ROC曲线下面积(AUC)达到0.877,提示miR-103b是糖尿病前期具有较高灵敏性和特异性的临床诊断标志物.生物信息学分析结果表明,hsa-miR-103b定位于人染色体5q34,并在10个物种间具有高度的保守性.靶基因预测共获得53个潜在的功能性靶向基因.进一步应用DAVID数据库和GO数据库进行靶基因功能富集分类,结果显示miR-103b靶基因功能主要涉及蛋白质氨基酸的磷酸化、RNA聚合酶II启动子转录调控和DNA依赖的转录调节,参与细胞周期、细胞生长增殖和细胞凋亡,调节功能蛋白结合并与PIP3结合激活下游信号通路.这些功能因子主要富集于MAPK、Wnt、胰岛素和Ras等信号通路,少量因子参与调节细胞周期和脂肪酸代谢等生理过程.结论:2型糖尿病前期患者血清中低水平miR-103b可能是潜在的2型糖尿病早期超前诊断标志物和治疗靶点.